Recombinant human fibronectin segment (rhFN1024) hydrogel carried hPDLSCs to repair diabetic trauma by activated NF-κB signaling pathway.

The accumulation of advanced glycation end products (AGEs) plays a crucial role in chronic inflammation and delayed wound healing in individuals with diabetes. In this context, fibronectin has been identified as a crucial protein that promotes the differentiation of human periodontal ligament stem cells (hPDLSCs) into myofibroblasts, which play a vital role in the repair of diabetic skin ulcers. This process is intimately associated with the integrin β1 receptor and the NF-κB signaling pathway, both crucial for cellular responses to fibronectin. To validate our hypothesis, we expressed rhFN₁₀₂₄, a recombinant protein containing the integrin β1 affinity-binding domain from human fibronectin segments 12-14. This protein was used to formulate a hydrogel for hPDLSCs. rhFN₁₀₂₄'s binding affinity to integrin β1 was confirmed by molecular docking and the cellular thermal shift assay (CETSA). We developed sh-ITGB1-hPDLSCs with stable ITGB1 knockdown using shRNA-ITGB1 and compared their proliferation, migration and adhesion to wild-type hPDLSCs. Morphological changes were observed via SEM, and α-SMA expression levels were measured in AGEs-damaged hPDLSCs. We created full-thickness wound models in diabetic mice to assess pharmacodynamics. The study showed that rhFN₁₀₂₄ stimulated hPDLSCs differentiation into myofibroblasts by boosting ITGB1 expression. rhFN₁₀₂₄ also reduced AGEs' negative effects on hPDLSCs, as seen through SEM analysis and α-SMA levels. In full-thickness wound models, hPDLSCs and rhFN₁₀₂₄ accelerated re-epithelialization and collagen synthesis. rhFN₁₀₂₄ is proposed to interact with the ITGB1 receptor on hPDLSCs, activating the NF-κB pathway to neutralize AGEs-induced pro-inflammatory cytokines. This study suggests rhFN₁₀₂₄ as a potential biomedical material for tissue repair.